Efficient generation of monoclonal antibodies from single human B cells by single cell RT-PCR and expression vector cloning.

@article{Tiller2008EfficientGO,
  title={Efficient generation of monoclonal antibodies from single human B cells by single cell RT-PCR and expression vector cloning.},
  author={Thomas Tiller and Eric Meffre and Sergey Yurasov and Makoto Tsuiji and Michel C. Nussenzweig and Hedda Wardemann},
  journal={Journal of immunological methods},
  year={2008},
  volume={329 1-2},
  pages={112-24}
}
We have developed an efficient strategy that combines immunoglobulin (Ig) gene repertoire analysis and Ig reactivity profiling at the single cell level. Based on surface marker expression individual cells at different stages of human B cell development are isolated by fluorescence-activated cell sorting. For each cell Ig heavy and corresponding Ig light chain gene transcripts are amplified by nested RT-PCR and cloned into eukaryotic expression vectors to produce monoclonal human antibodies of… CONTINUE READING
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